# Incubate on bench overnight or until water is about room temperature (~4 hr). At the end of this process, the complimentary ssDNA's are hybridized into dsDNA linker at 1mM final concentration.

+

# Incubate on bench overnight or until water is about room temperature (~4 hr). At the end of this process, the complimentary ssDNA's are hybridized into as single dsDNA linker with sticky ends at 1mM final concentration.

-

# Dilute the 1mM linker 1/10 (~10 μL 1mM linker into ~ 90 μL H<sub>2</sub>O, may need to be adjusted if some volume is lost and, for example only 9 μL 1mM linker is recovered). Mix and spin.

+

# Perform two 1/100 dilutions down to 100 nM (label all tubes clearly):

-

# Continue 1/10 dilutions down to 100 nM (label all tubes clearly):

+

#* 2 μL 1mM linker + 198 μL H<sub>2</sub>O = 200 μL 10μM linker

-

#* 20 μL 100μM linker + 180 μL H<sub>2</sub>O = 200 μL 10μM linker

+

#* 2 μL 10μM linker + 198 μL H<sub>2</sub>O = 200 μL 100nM linker

-

#* 20 μL 10μM linker + 180 μL H<sub>2</sub>O = 200 μL 1μM linker

+

# Store 1mM, 10μM, and 100nM stocks at -40 °C in latest "Primer" box. Keep an aliquot of the 100nM linker in your own box for use in constructions.

Incubate on bench overnight or until water is about room temperature (~4 hr). At the end of this process, the complimentary ssDNA's are hybridized into as single dsDNA linker with sticky ends at 1mM final concentration.

Perform two 1/100 dilutions down to 100 nM (label all tubes clearly):

2 μL 1mM linker + 198 μL H2O = 200 μL 10μM linker

2 μL 10μM linker + 198 μL H2O = 200 μL 100nM linker

Store 1mM, 10μM, and 100nM stocks at -40 °C in latest "Primer" box. Keep an aliquot of the 100nM linker in your own box for use in constructions.

Notes

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